Stimulatory effect of insecticides on partially purified P-glycoprotein ATPase from the resistant pest Helicoverpa armigera
Stimulatory effect of insecticides on partially purified P-glycoprotein ATPase from the resistant pest Helicoverpa armigera
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DOI：
10.1139/o06-194
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发表时间：
2006-12-01
影响因子：
2.9
通讯作者：
Sreeramulu, Kuruba
中科院分区：
文献类型：
--
作者：
Aurade, Ravindra;Jayalakshmi, Senigala K.;Sreeramulu, Kuruba
使用针对核苷酸结合域中表位的 C219 抗体，在抗杀虫剂害虫棉铃虫（鳞翅目：NoctUidae）的膜制备物中检测到 P-糖蛋白样蛋白 (Ha-Pgp)。该蛋白质经过部分纯化，发现是一种具有 ATP 酶活性的糖蛋白。 SDS-PAGE证实高分子量糖蛋白（150 kDa）在抗性害虫中过度表达，但在易感害虫中未检测到。将部分纯化的 Ha-Pgp ATP 酶重构为脂蛋白体，发现一些杀虫剂，即久效磷、硫丹、氯氰菊酯、氰戊菊酯和甲基对硫磷，可刺激 ATP 酶活性。各种抑制剂对部分纯化的Ha-Pgp的影响表明，原钒酸盐是其ATP酶活性的有效抑制剂，在2 mmol/L浓度下可抑制90%。其他抑制剂，如 EDTA、叠氮化钠和钼酸盐，仅导致活性降低 20%。 Ha-Pgp 的结构和功能细节对于制定克服该害虫杀虫剂抗性的策略非常重要。
A P-glycoprotein-like protein (Ha-Pgp) was detected in a membrane preparation from the insecticide-resistant pest Helicoverva armigera (Lepidoptera: NoctUidae) using C219 antibodies that are directed towards an epitope in the nucleotide-binding domains. This protein was partially purified and found to be a glycoprotein displaying ATPase activity. SDS-PAGE confirmed that a high molecular mass glycoprotein (150 kDa) was overexpressed in resistant pests, but was not detected in susceptible pests. The partially purified Ha-Pgp ATPase was reconstituted into proteoliposomes and it was found that some insecticides, namely, monocrotophos, endosulfan, cypermethrin, fenvalerate, and methylparathion, stimulated the ATPase activity. The effect of various inhibitors on partially purified Ha-Pgp showed that orthovanadate is a potent inhibitor of its ATPase activity, inhibiting it by 90% at a concentration of 2 mmol/L. Other inhibitors, such as EDTA, sodium azide, and molybdate resulted in only a 20% decrease in activity. Details of the structure and function of Ha-Pgp will be important in the development of strategies to overcome insecticide resistance in this pest.